基于小孢子培养的芥蓝单倍体育种技术研究

    Study on Microspore Culture-Based Haploid Breeding Technology of Chinese Kale(Brassica oleracea)

    • 摘要: 【目的】为提高芥蓝育种效率,对基于小孢子培养的芥蓝单倍体育种技术流程进行研究。【方法】利用离心管进行机械破碎替代手工研磨提取芥蓝游离小孢子,采用秋水仙素处理芥蓝再生苗,并对其他技术环节进行优化。【结果】机械破碎提取芥蓝小孢子的胚状体诱导率(11.96%)与手工研磨的胚状体诱导率(10.80%)无显著差异,可用于批量处理样品、提高芥蓝小孢子提取效率;两种方法采集芥蓝小孢子进行培养,共诱导产生胚状体 255 个,经继代、生根培养获得 112 个芥蓝再生株系;再生植株大多为单倍体,秋水仙素处理使其存活率和染色体加倍频率提高;再生株系单株自交获得芥蓝 DH 系 41 个,其中秋水仙素处理的 DH 成系率达56.45%,是未处理的 4.07 倍。【结论】建立了一套较完整有效的芥蓝单倍体育种技术流程,可在芥蓝育种实践中有推广应用,提高芥蓝育种效率。

       

      Abstract: 【Objective】To improve the breeding efficacy of Chinese kale, the procedure of microspore culture-based haploid breeding technology was studied.【Method】Mechanical crushing with centrifuge tube was used to extract isolated microspores of Chinese kale instead of artificial grinding, regenerated plantlets were treated with cholchicine, and other steps were also optimized.【Result】There was no significant difference in the embryoid induction rate(11.96%)of microspores of Chinese kale extracted by mechanical crushing and that by artificial grinding(10.80%). Mechanical crushing could be used to treat samples in a batch to improve the extraction efficiency of microspores of Chinese kale. The microspores of Chinese kale collected by the two methods were cultured, a total of 255 embryoids were induced and 112 regenerated lines obtained after subculture and rooting culture. The treatment with colchicine improved chromosome doubling of regeneration plantlets(most of which were haploid), leading to a DH attainment rate of 56.45%, 3.07 times higher than that of non treatment. Through the procedure, 41 DH lines were obtained from different donor varieties.【Conclusion】An integrated and effective microspore culture-based haploid breeding procedure was developed, which could be applied in breeding practice of Chinese kale to improve breeding efficacy.

       

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